OALib Journal期刊
ISSN: 2333-9721
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毛细管凝胶电泳紫外检测核酸灵敏度
, PP. 1161-1166
Keywords: 脱氧核糖核酸,毛细管电泳,基质场放大,柱头场放大,聚合酶链式反应产物
Abstract:
以已知浓度的DNAMarker为标准样品,常规压力进样,电动进样,柱头场放大及联合使用基质场放大和柱头场放大等方法进行CGE-UV分析,对比不同进样方法的检测灵敏度。以聚合酶链式反应(PCR)后的高盐DNA样品验证方法的可靠性。当DNA样品稀释达40万倍,与电动进样和压力进样相比,在对峰形和峰宽无明显影响下,将电动进样的时间延至420s,三羟甲基氨基甲烷-盐酸缓冲液(TE)浓度降低至1.5%,柱头场放大灵敏度分别提高了约28和90倍;联合使用基质场放大和柱头场放大后,灵敏度分别提高3760和12000倍。DNA的检出限达0.1μg/L(S/N=3,CGE-FASI-UV)。同时以本法分析PCR后的DNA产物获得了极高的灵敏度(分别比普通的压力进样和电动进样提高50477倍和33354倍)。本实验采用基质场放大和柱头场放大联用,方法操作简单,灵敏度高,适用于高盐微量的DNA样品分析。
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